Establishment of a simple and efficient platform for car-t cell generation and expansion: from lentiviral production to in vivo studies

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Picanco-Castro, Virginia ; Moco, Pablo Diego ; Mizukami, Amanda ; Vaz, Leticia Delfini ; Fernandes Pereira, Marcelo de Souza ; Silvestre, Renata Nacasaki ; Cottas de Azevedo, Julia Teixeira ; Bomfim, Aline de Sousa ; de Abreu Neto, Mario Soares ; Ribeiro Malmegrim, Kelen Cristina ; Swiech, Kamilla ; Covas, Dimas Tadeu

• Document type: Journal article
• Source: Hematology, Transfusion and Cell Therapy; v. 42, n. 2, p. 9-pg., 2020-04-01.

Abstract

Introduction: Adoptive transfer of T cells expressing a CD19-specific chimeric antigen receptor (CAR) has shown impressive response rates for the treatment of CD19 + B-cell malignancies in numerous clinical trials. The CAR molecule, which recognizes cell-surface tumor-associated antigen independently of human leukocyte antigen (HLA), is composed by one or more signaling molecules to activate genetically modified T cells for killing, proliferation, and cytokine production. Objectives: In order to make this treatment available for a larger number of patients, we developed a simple and efficient platform to generate and expand CAR-T cells. Methods: Our approach is based on a lentiviral vector composed by a second-generation CAR that signals through a 41BB and CD3-zeta endodomain. Conclusions: In this work, we show a high-level production of the lentiviral vector, which was successfully used to generate CAR-T cells. The CAR-T cells produced were highly cytotoxic and specific against CD19+ cells in vitro and in vivo, being able to fully control disease progression in a xenograft B-cell lymphoma mouse model. Our work demonstrates the feasibility of producing CAR-T cells in an academic context and can serve as a paradigm for similar institutions. Nevertheless, the results presented may contribute favoring the translation of the research to the clinical practice. (C) 2019 Associacao Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. (AU)

• FAPESP's process: 12/23228-4 - In vitro expansion of mesenchymal stromal cells and secretome characterization: therapeutic and biotechnological applications
• Grantee: Amanda Mizukami Martins
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/09491-8 - Murine Leukemia/Lymphoid Lymphoma models for evaluating antitumor effects of CAR-CD19-modified T cells
• Grantee: Marcelo de Souza Fernandes Pereira
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 16/08374-5 - Modification of T cells with anti-CD19 chimeric antigen receptor
• Grantee: Pablo Diego Moço
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC